Experimental quantification of annular dark-field images in scanning transmission electron microscopy

This paper reports on a method to obtain atomic resolution Z-contrast (high-angle annular dark-field) images with intensities normalized to the incident beam. The procedure bypasses the built-in signal processing hardware of the microscope to obtain the large dynamic range necessary for consecutive measurements of the incident beam and the intensities in the Z-contrast image. The method is also used to characterize the response of the annular dark-field detector output, including conditions that avoid saturation and result in a linear relationship between the electron flux reaching the detector and its output. We also characterize the uniformity of the detector response across its entire area and determine its size and shape, which are needed as input for image simulations. We present normalized intensity images of a SrTiO(3) single crystal as a function of thickness. Averaged, normalized atom column intensities and the background intensity are extracted from these images. The results from the approach developed here can be used for direct, quantitative comparisons with image simulations without any need for scaling.     

Advances in TEM orientation microscopy by combination of dark-field conical scanning and improved image matching

A new approach to automatic TEM‐based orientation microscopy is presented, which is based on a combination of the techniques of dark-field conical scanning and improved image matching, and a diffraction pattern simulation method. For indexing, a full experimental diffraction pattern is compared to all possible pre-calculated diffraction patterns for the given structure by image matching. In order to speed up this relatively calculation-intensive algorithm, polar transformation and, most important, circular projection that increase the speed of pattern indexing by a factor of about 50 are proposed. A microstructure of submicron scale and crystallographic orientations in nanocrystalline materials are measured successfully. It is proposed that the taken approach of dark-field conical scanning and improved image matching may be, in principle, better suited for TEM-based orientation microscopy than serial orientation mapping.     

Three-dimensional imaging in double aberration-corrected scanning confocal electron microscopy, part I: elastic scattering

A transmission electron microscope fitted with both pre-specimen and post-specimen spherical aberration correctors enables the possibility of aberration-corrected scanning confocal electron microscopy. Imaging modes available in this configuration can make use of either elastically or inelastically scattered electrons. In this paper we consider image contrast for elastically scattered electrons. It is shown that there is no linear phase contrast in the confocal condition, leading to very low contrast for a single atom. Multislice simulations of a thicker crystalline sample show that sample vertical location and thickness can be accurately determined. However, buried impurity layers do not give strong, nor readily interpretable contrast. The accompanying paper examines the detection of inelastically scattered electrons in the confocal geometry.     

Bloch wave-based calculation of imaging properties of high-resolution scanning confocal electron microscopy

An efficient, Bloch wave-based method is presented for simulation of high-resolution scanning confocal electron microscopy (SCEM) images. The latter are predicted to have coherent nature, i.e. to exhibit atomic contrast reversals depending on the lens defocus settings and sample thickness. The optimal defocus settings are suggested and the 3D imaging capabilities of SCEM are analyzed in detail. In particular, by monitoring average image intensity as a function of the probe focus depth, it should be possible to accurately measure the depth of a heavy-atom layer embedded in a light-element matrix.     

Three-dimensional imaging in double aberration-corrected scanning confocal electron microscopy, part II: inelastic scattering

The implementation of spherical aberration-corrected pre- and post-specimen lenses in the same instrument has facilitated the creation of sub-Angstrom electron probes and has made aberration-corrected scanning confocal electron microscopy (SCEM) possible. Further to the discussion of elastic SCEM imaging in our previous paper, we show that by performing a 3D raster scan through a crystalline sample using inelastic SCEM imaging it will be possible to determine the location of isolated impurity atoms embedded within a bulk matrix. In particular, the use of electron energy loss spectroscopy based on inner-shell ionization to uniquely identify these atoms is explored. Comparisons with scanning transmission electron microscopy (STEM) are made showing that SCEM will improve both the lateral and depth resolution relative to STEM. In particular, the expected poor resolution of STEM depth sectioning for extended objects is overcome in the SCEM geometry.     

Imaging of high-angle annular dark-field scanning transmission electron microscopy and observations of GaN-based violet laser diodes

The first part of this paper is devoted to physics, to explain high‐angle annular dark‐field scanning transmission electron microscopy (HAADF‐STEM) imaging and to interpret why HAADF‐STEM imaging is incoherent, instructing a strict definition of interference and coherence of electron waves. Next, we present our recent investigations of InGaN/GaN multiple quantum wells and AlGaN/GaN strained‐layer superlattice claddings in GaN‐based violet laser diodes, which have been performed by HAADF‐STEM and high‐resolution field‐emission gun scanning electron microscopy.     

The impact of surface and retardation losses on valence electron energy-loss spectroscopy

The inelastic scattering of fast electrons transmitting thin foils of silicon (Si), silicon nitride (Si(3)N(4)), gallium arsenide (GaAs), gallium nitride (GaN) and cadmium selenide (CdSe) was analyzed using dielectric theory. In particular, the impact of surface and bulk retardation losses on valence electron energy-loss spectroscopy (VEELS) was studied as a function of the foil thickness. It is shown that for the materials analyzed, surface and retardation losses can cause a systematic, thickness-dependent modulation of the dielectric volume losses, which can hamper the determination of the bulk dielectric data as well as the identification of band-gap and interband transition energies by VEELS. For Si and GaAs, where the dielectric function is strongly peaked with high absolute values, retardation losses lead to additional intensity maxima in the spectrum. For thin films of these materials (below approximately 100 nm), the additional intensity maxima are related to retardation effects due to the finite size of the sample leading to the excitation of guided light modes. For thicker films, exceeding about 200 nm, the intensity maxima are caused by bulk retardation losses, i.e., Cerenkov losses. Although thickness-dependent modulations were observed for Si(3)N(4), GaN and CdSe, the form of the dielectric functions and their lower maxima, means that for TEM samples < 100 nm thick, the band-gap energies of these materials can be accurately identified by VEELS. Guidelines are given that allow for forecasting the impact of surface and retardation losses on VEELS.     

Practical factors affecting the performance of a thin-film phase plate for transmission electron microscopy

A number of practical issues must be addressed when using thin carbon films as quarter-wave plates for Zernike phase-contrast electron microscopy. We describe, for example, how we meet the more stringent requirements that must be satisfied for beam alignment in this imaging mode. In addition we address the concern that one might have regarding the loss of some of the scattered electrons as they pass through such a phase plate. We show that two easily measured parameters, (1) the low-resolution image contrast produced in cryo-EM images of tobacco mosaic virus particles and (2) the fall-off of the envelope function at high resolution, can be used to quantitatively compare the data quality for Zernike phase-contrast images and for defocused bright-field images. We describe how we prepare carbon-film phase plates that are initially free of charging or other effects that degrade image quality. We emphasize, however, that even though the buildup of hydrocarbon contamination can be avoided by heating the phase plates during use, their performance nevertheless deteriorates over the time scale of days to weeks, thus requiring their frequent replacement in order to maintain optimal performance.     

The influence of relativistic energy losses on bandgap determination using valence EELS

Since monochromated transmission electron microscopes have become available, the determination of bandgaps and optical properties using electron energy loss spectrometry (EELS) has again attracted interest. The underlying idea is very simple: below the bandgap energy no transitions can contribute to the valence EELS signal. However, the bandgap cannot be directly read out from the recorded data. Therefore the optical properties cannot be determined correctly from the low loss using the Kramers-Kronig relations. We will discuss under which conditions relativistic effects may be suppressed. It is demonstrated that scanning TEM (STEM) geometry is not applicable for most bandgap measurements.     

Charge compensation by in-situ heating for insulating ceramics in scanning electron microscope

With a steady temperature increase under high vacuum (HV) in an environmental scanning electronic microscope, we observed charge-free characterization and fine secondary electron (SE) images in focus for insulating ceramics (alumina (Al₂O₃), aluminum nitride (AlN), pure magnesium silicate (Mg₂SiO₄)). The sample current I_sc increased from −8.18×10⁻¹³ to 2.76×10⁻⁷ A for Al₂O₃ and −9.28×10⁻¹² to 2.77×10⁻⁶ A for AlN with the temperature increased from 298 to 633 K. The surface conductance σ increased from 5.6×10⁻¹³ to 5.0×10⁻¹¹/Ω for Al₂O₃ and 1.1×10⁻¹² to 1.0×10⁻⁷/Ω for AlN with the temperature increased from 363 to 593 K. The SE image contrast obtained via heating approach in high vacuum with an Everhart–Thornley SE-detector was better than that via conventional approach of electron–ion neutralization in low vacuum (LV) with a gaseous SE-detector. The differences of compensation temperatures for charge effects indicate dielectric and thermal properties, and band structures of insulators. The charge compensation mechanisms of heating approach mainly relate to accelerated release of trapped electrons on insulating surface and to increase of electron emission yield by heating.     

A systematic approach to choosing parameters for modelling fine structure in electron energy-loss spectroscopy

A potential methodology is presented for the systematic prediction of EELS edges using DFT, suitable for codes that calculate ELNES for a specific atom in a unit cell. The method begins with the selection of a unit cell, chosen as the smallest cell that still provides a physically valid representation of the bulk material. Within this small cell, a single electron core–hole is included in the atom for which the EELS ionisation edge is to be calculated. The basis-set size and k-point mesh of the DFT calculation are converged specifically against the predicted EELS result. Subsequently, the cell size is increased until the theoretical core–holes no longer interfere. At this point one can then modify the exact core–hole approximation. This methodology was applied to the new EELS module of the CASTEP pseudopotential DFT code, as well as the all-electron code Wien2k. Aluminium K edges were investigated for various aluminium metal systems. It was observed that as the cell size was increased the predicted EELS result became less sensitive to the exact core–hole approximation used. It was noted however that due to high screening in metals a ground state single cell calculation is often acceptable. The semiconductor aluminium nitride (wurtzite form) was also investigated. It was observed that for both Wien2k and CASTEP, with careful convergence of the key DFT code parameters, single cell ground state calculations gave a reasonable agreement with experiment, contrary to what might be expected for a semiconductor with a large band gap. This was particularly true of the Wien2k result. Given the greater computational effort required for supercell calculations, these results are likely to form the beginnings of a detailed investigation into accepted methods of ELNES predictions.     

“Ab initio” structure solution from electron diffraction data obtained by a combination of automated diffraction tomography and precession technique

Using a combination of our recently developed automated diffraction tomography (ADT) module with precession electron technique (PED), quasi-kinematical 3D diffraction data sets of an inorganic salt (BaSO₄) were collected. The lattice cell parameters and their orientation within the data sets were found automatically. The extracted intensities were used for “ab initio” structure analysis by direct methods. The data set covered almost the complete set of possible symmetrically equivalent reflections for an orthorhombic structure. The structure solution in one step delivered all heavy (Ba, S) as well as light atoms (O). Results of the structure solution using direct methods, charge flipping and maximum entropy algorithms as well as structure refinement for three different 3D electron diffraction data sets were presented.     

Depth sectioning in scanning transmission electron microscopy based on core-loss spectroscopy

Recent and ongoing improvements in aberration correction have opened up the possibility of depth sectioning samples using the scanning transmission electron microscope in a fashion similar to the confocal scanning optical microscope. We explore questions of principle relating to image interpretability in the depth sectioning of samples using electron energy loss spectroscopy. We show that provided electron microscope probes are sufficiently fine and detector collection semi-angles are sufficiently large we can expect to locate dopant atoms inside a crystal. Furthermore, unlike high angle annular dark field imaging, electron energy loss spectroscopy can resolve dopants of smaller atomic mass than the supporting crystalline matrix.     

Visualization of magnetic domains by near-field scanning microwave microscope

A near-field scanning microwave microscope (NSMM) system was used for the investigation of magnetic properties of a hard disk (HD) under an external magnetic field. To demonstrate local microwave characterization of magnetic domains by NSMM, we scanned the HD surface by measuring the microwave reflection coefficient S₁₁ of the NSMM at an operating frequency near 4.4 GHz. The NSMM offers a reliable means for quantitative measurement of magnetic domains with high spatial resolution and sensitivity.     

In situ measurements and transmission electron microscopy of carbon nanotube field-effect transistors

We present the design and operation of a transmission electron microscopy (TEM)-compatible carbon nanotube (CNT) field-effect transistor (FET). The device is configured with microfabricated slits, which allows direct observation of CNTs in a FET using TEM and measurement of electrical transport while inside the TEM. As demonstrations of the device architecture, two examples are presented. The first example is an in situ electrical transport measurement of a bundle of carbon nanotubes. The second example is a study of electron beam radiation effect on CNT bundles using a 200 keV electron beam. In situ electrical transport measurement during the beam irradiation shows a signature of wall- or tube-breakdown. Stepwise current drops were observed when a high intensity electron beam was used to cut individual CNT bundles in a device with multiple bundles.     

Determination of chromium valence over the range Cr(0)–Cr(VI) by electron energy loss spectroscopy

Chromium is a redox active 3d transition metal with a wide range of valences (−2 to +6) that control the geochemistry and toxicity of the element. Therefore, techniques that measure Cr valence are important bio/geochemical tools. Until now, all established methods to determine Cr valence were bulk techniques with many specific to a single, or at best, only a few oxidation state(s). We report an electron energy loss spectroscopy (EELS) technique along with an extensive suite of affined reference spectra that together, unlike other methods, can determine Cr valence (or at least constrain the possible valences) at high-spatial resolution (tens-of-nanometer scale) across a wide valence range, Cr(0)–Cr(VI). Fine structure of Cr-L_2,3 edges was parametrized by measurement of the chemical shift of the L₃ edge and the ratio of integrated intensity under the L₃ and L₂ edges. These two parameterizations were correlated to Cr valence and also the dⁿ orbital configuration which has a large influence on L-edge fine structure. We demonstrate that it is not possible to unambiguously determine Cr valence from only one fine-structure parameterization which is the method employed to determine metal valence by nearly all previous EELS studies. Rather, multiple fine-structure parameterizations must be used together if the full range of possible Cr valences is considered. However even with two parameterizations, there are limitations. For example, distinguishing Cr(IV) from Cr(III) is problematic and it may be difficult to distinguish low-spin Cr(II) from Cr(III). Nevertheless, when Cr is known to be divalent, low- and high-spin dⁿ orbital configurations can be readily distinguished.     

Morphological investigations of cells that adhered to the irregular patterned polydimethylsiloxane (PDMS) surface without reagents

Polydimethylsiloxane (PDMS) surface consisting irregular pattern was investigated to develop cell-based biochip using PDMS. PDMS surface was modified with nano- and micro-combined patterns using surface deformation technology. Hydrophobicity of nano-patterned PDMS surface was sustained. Nevertheless it has irregular patterns consisting of micro- and nano-patterns. According to atomic force microscopy (AFM), scanning electron microscopy (SEM) and confocal microscopy results by immunostaining method, human mammary epithelial cells (HMEC) adhered well on irregularly patterned surface without any reagents such as gelatin and collagen, compared to commercial culture dish. It implies PDMS material can be utilized as template for cell-based biochip without any reagents.     

Nanoscale film formation of ferritin and its application to biomemory device

A redox protein, ferritin is used as a functional constituent of the developed biomemory device. The concept of molecular device mainly depends on the solidification of biomolecules of interest and on the realization of properties of molecule immobilized on a selected substrate. Here, we immobilized the biomolecule, ferritin protein on gold substrate using an organic linker 11-mercaptoundecanoic acid (11-MUA). The immobilization of the protein on the gold substrate was confirmed by surface plasmon spectroscopy, Raman spectroscopy, and atomic force microscopy (AFM). The basic two memory functions, reading and writing of the developed biomemory device, were investigated by open-circuit potential amperometry (OCPA) using the redox property of the biomolecule of interest. The surface topography investigation by scanning tunneling microscopy (STM) shows that the robustness of the ferritin-based biomemory device was validated by the repeated electrochemical performance. These results show the developed biomemory device as a step towards the protein-based nanobiochip.     

The statistics of the thermal motion of the atoms during imaging process in transmission electron microscopy and related techniques

The concern of this work is the influence of the thermal motion of the atoms on electron scattering simulations, used for quantitative interpretation of results in high-resolution electron microscopy. We distinguish between the influence of inelastic phonon excitation and the effect of a moving lattice on images generated by elastically scattered electrons. It is shown that, analog to aberrations, the impact of a moving lattice differs substantially with respect to different imaging conditions and cannot be described by the Debye–Waller damping applicable in XRD. We derive a new formalism, based on the frozen lattice and multislice approach, to incorporate the statistics of the thermal motion into elastic TEM imaging simulations, taking into account different imaging conditions. The averaging over different atom positions is generally performed within a density matrix framework, which can be linearized in the special case of off-axis electron holography. All findings are supported by explicit numerical simulations: molecular dynamics simulations are performed to get a realistic thermal motion and the electron scattering simulations are performed within the new multislice algorithm.